Confidence intervals in procedural dermatology: an intuitive approach to interpreting data.
Significant differences observed in therapeutic trials in procedural dermatology are typically denoted by p values of less than .05. Alternatively, significance can be conveyed by use of confidence intervals. The purpose of this article is to clarify how confidence intervals convey the same information about outcomes as p values, albeit in a slightly different manner. (1) Selective review of textbooks and other relevant literature and (2) presentation of a brief tutorial describing confidence interval determination for therapeutic clinical trials comparing differences between means of two groups. Routine use of confidence intervals is an intuitively satisfying means for conveying the statistical significance of results and can be used in combination with p values for understanding these results. Specifically, confidence intervals are a useful tool for indicating the size, spread, and direction of the observed differences. Unfortunately, dermatologic surgery trials tend to have low sample sizes, which frequently result in outcomes below the threshold of statistical significance (p > .05, or confidence intervals including 1.00). In the absence of statistical significance, neither p values nor confidence intervals yield definitive results. Confidence intervals can complement p values as a means for explaining statistical significant differences. When differences are not statistically significant but are clinically significant and approach statistical significance, neither p values nor confidence intervals can definitively establish whether the observed trends are indicative of an underlying difference. In these cases, common in procedural dermatology, larger, better designed, randomized prospective trials are needed.


The individualistic fallacy, ecological studies and instrumental variables: a causal interpretation.
The validity of ecological studies in epidemiology for inferring causal relationships has been widely challenged as observed associations could be biased by the Ecological Fallacy. We reconsider the important design components of ecological studies, and discuss the conditions that may lead to spurious associations. Ecological associations are useful and valid when the ecological exposures can be interpreted as Instrumental Variables. A suitable example may be a time series analysis of environmental pollution (e.g. particulate matter with an aerodynamic diameter of <10 micrometres; PM10) and health outcomes (e.g. hospital admissions for acute myocardial infarction) as environmental pollution levels are a cause of individual exposure levels and not just an aggregate measurement. Ecological exposures may also be employed in situations (perhaps rare) where individual exposures are known but their associations with health outcomes are confounded by unknown or unquantifiable factors. Ecological associations have a notorious reputation in epidemiology and individualistic associations are considered superior to ecological associations because of the "ecological fallacy". We have argued that this is incorrect in situations in which ecological or aggregate exposures can serve as an instrumental variable and associations between individual exposure and outcome are likely to be confounded by unmeasured variables.


Effects of secondary binding by activator and inhibitor peptides on covalent intermediates of pig pepsin.
A number of peptides were found to increase the activity of pig pepsin towards small synthetic substrates. The activators increase transpeptidation of both the acyl-transfer and the amino-transfer types by as much as 45-fold. The effect on hydrolysis varies from inhibition to modest activation, but is always less than the effect on transpeptidation. The kinetics of substrate cleavage are the converse of non-competitive inhibition and show an increase in kcat. and no effect on Km values. Lineweaver-Burk plots of results obtained in the presence of the activators indicate a substrate activation at high substrate concentration. This appears to be a co-operative effect, since it is not observed in the absence of the activators. The activation is greatest at pH 4.7, less at pH 3.4, and at pH 2.0 is observable only with some of the activator peptides. The results show directly the effect of secondary binding on the catalytic efficiency of pepsin. The most effective activators are those that are most hydrophobic. The results suggest that binding in the secondary binding sites causes an increase in hydrophobicity in the catalytic site which results in increased stability of the acyl and amino intermediates, and preferential reaction with acceptors other than water. The implication that the present results strengthen the case for a role of covalent intermediates in the hydrolysis of good substrates (high kcat. values) is discussed.


Concordance between administrative health data and medical records for diabetes status in coronary heart disease patients: a retrospective linked data study.
Administrative data are a valuable source of estimates of diabetes prevalence for groups such as coronary heart disease (CHD) patients. The primary aim of this study was to measure concordance between medical records and linked administrative health data for recording diabetes in CHD patients, and to assess temporal differences in concordance. Secondary aims were to determine the optimal lookback period for identifying diabetes in this patient group, whether concordance differed for Indigenous people, and to identify predictors of false positives and negatives in administrative data. A population representative sample of 3943 CHD patients hospitalized in Western Australia in 1998 and 2002-04 were selected, and designated according to the International Classification of Diseases (ICD) version in use at the time (ICD-9 and ICD-10 respectively). Crude prevalence and concordance were compared for the two samples. Concordance measures were estimated from administrative data comparing diabetes status recorded on the selected CHD admission ('index admission') and on any hospitalization in the previous 1, 2, 5, 10 or 15 years, against hospital medical records. Potential modifiers of agreement were determined using chi-square tests and multivariable logistic regression models. Identification of diabetes on the index CHD admission was underestimated more in the ICD-10 than ICD-9 sample (sensitivity 81.5% versus 91.1%, underestimation 15.1% versus 4.4% respectively). Sensitivity increased to 89.6% in the ICD-10 period using at least 10 years of hospitalization history. Sensitivity was higher and specificity lower in Indigenous patients, and followed a similar pattern of improving concordance with increasing lookback period. Characteristics associated with false negatives for diabetes on the index CHD hospital admission were elective admission, in-hospital death, principal diagnosis, and in the ICD-10 period only, fewer recorded comorbidities. The accuracy of identifying diabetes status in CHD patients is improved in linked administrative health data by using at least 10 years of hospitalization history. Use of this method would reduce bias when measuring temporal trends in diabetes prevalence in this patient group. Concordance measures are as reliable in Indigenous as non-Indigenous patients.


Ageing doctors.
Doctors are neither more nor less susceptible than the general population to the effects of ageing. The relevance of deterioration with age depends on the nature of the work undertaken. Reduced muscle strength and visual and auditory deterioration can compromise clinical ability. Accumulation of chronic disease further reduces capacity. Cognitive decline is of particular importance, as good medical care requires considerable cognitive function. Patient safety is paramount, yet older doctors are an important part of the medical workforce and their value should be recognised. Changes in patient case mix, work place support systems and individual adjustments can assist safe practice. Deterioration in health should be acknowledged and requires proactive management. Current methods of ensuring competence are inadequate for supporting ageing doctors. A new initiative is recommended comprising collaboration between regulators, colleges and employing institutions to support the ageing doctor in providing safe and effective practice.


The effects of surgical stress and short-term fasting on protein synthesis in vivo in diverse tissues of the mature rat.
1. We measured fractional rates of protein synthesis, capacities for protein synthesis (i.e. RNA/protein ratio) and efficiencies of protein synthesis (i.e. protein-synthesis rate relative to RNA content) in fasted (24 or 48 h) or fasted/surgically stressed female adult rats. 2. Of the 15 tissues studied, fasting caused decreases in protein content in the liver, gastrointestinal tract, heart, spleen and tibia. There was no detectable decrease in the protein content of the skeletal muscles studied. 3. Fractional rates of synthesis were not uniformly decreased by fasting. Rates in striated muscles, uterus, liver, spleen and tibia were consistently decreased, but decreases in other tissues (lung, gastrointestinal tract, kidney or brain) were inconsistent or not detectable, suggesting that, in many tissues in the mature rat, protein synthesis was not especially sensitive to fasting. 4. In fasting, the decreases in fractional synthesis rate resulted from changes in efficiency (liver and tibia) or from changes in efficiency and capacity (heart, diaphragm, plantaris and gastrocnemius). In the soleus, the main change was a decrease in capacity. 5. Surgical stress increased fractional rates of protein synthesis in diaphragm (where there were increases in both efficiency and capacity) by about 50%, in liver by about 20%, in spleen by about 40%, and possibly also in the heart. In liver and spleen, capacities were increased. In other tissues (including the skeletal muscles), the fractional rates of protein synthesis were unaffected by surgical stress.


Preventive child health care at elementary school age: The costs of routine assessments with a triage approach.
Triage in Preventive Child Health Care (PCH) assessments could further the efficient use of human resources and budgets and therefore make extra care possible for children with specific needs. We assessed the costs of routine PCH assessments with and without triage for children aged 5/6 years and 10/11 years. In a triage approach, PCH assistants conduct pre-assessments to identify children requiring follow-up assessments by a physician or nurse. In the usual approach, all children are assessed by a physician and an assistant (children aged 5/6 years) or a nurse (children aged 10/11 years). All the direct costs of conducting routine PCH assessments with the triage and usual approach were assessed using a bottom-up micro-costing approach. In four PCH services in the Netherlands, two using triage and two the usual approach, professionals completed questionnaires about time spent on assessments, including time related to non-attendance at assessments, the referral of children and administration. The projected costs for PCH professionals working on PCH assessments amounted to €5.2 million per cohort of 100,000 children aged 5/6 years in the triage approach, and €7.6 million in the usual approach. The projected costs in both approaches for children aged 10/11 years were about €4 million per 100,000 children. The triage approach to PCH resulted in a projected cost reduction of about one-third, compared with usual practice, for routine assessments by physicians of children aged 5/6 years. There are minimal cost savings in the group of children aged 10/11 years when nurses are involved and so other considerations such as workforce shortages would be required to justify a change to a triage approach. Further research is needed to investigate the differences in costs of care after the completion of the routine assessments.


Predictors of parenting concern in a Mother and Baby Unit over a 10-year period.
To review the characteristics of 462 mothers consecutively admitted to a co-joined mother and baby unit and to examine the incidence and predictors of social services input and separate discharge. Admissions to the Birmingham Mother and Baby Unit from 1998 to 2007 were reviewed. Demographic and clinical information, social services involvement and parenting outcome data were collected from case notes and computerized records. One hundred and forty six (146) women received some form of social services input (32%); 44 cases of separate discharge were identified (10%). Diagnosis, involuntary admission, single status, younger age, Afro-Caribbean ethnicity and social services referral predicted parenting concern. Only 9% of separations were to women suffering a new postpartum mental illness; most were to women experiencing the continuation of an existing illness. Poor parenting skills, absence of a good partner relationship and alcohol/substance misuse were frequently reported in those discharged without their baby. The incidence of separate discharge is lower than in other studies. Potential reasons for this difference are discussed. Further work must be conducted to understand the social inequities and regional differences in social service intervention during perinatal psychiatric admission and determine if parenting outcomes can be improved in women at risk of separation.


Simple and efficient generation of virus-specific T cells for adoptive therapy using anti-4-1BB antibody.
Although recent studies of virus-specific T-cell (VST) therapy for viral infections after allogeneic hematopoietic stem cell transplantation have shown promising results, simple and less time-intensive and labor-intensive methods are required to generate VSTs for the wider application of VST therapy. We investigated the efficacy of anti-CD28 and anti-4-1BB antibodies, which can provide T cells with costimulatory signals similar in strength to those of antigen-presenting cells, in generating VSTs. When peripheral blood mononuclear cells were stimulated with viral peptides together with isotype control, anti-CD28, or anti-4-1BB antibodies, anti-4-1BB antibodies yielded the highest numbers of VSTs, which were on an average 7.9 times higher than those generated with isotype control antibody. The combination of anti-CD28 and anti-4-1BB antibodies did not result in increased numbers of VSTs compared with anti-4-1BB antibody alone. Importantly, the positive effect of anti-4-1BB antibody was observed regardless of the epitopes of the VSTs. In contrast, the capacity of dendritic cells (DCs) to generate VSTs differed considerably depending on the epitopes of the VSTs. Furthermore, the numbers of VSTs generated with DCs were at most similar to those generated with the anti-4-1BB antibody. Generation of VSTs with anti-4-1BB antibody did not result in excessive differentiation or deteriorated function of the generated VSTs compared with those generated with control antibody or DCs. In conclusion, VSTs can be generated rapidly and efficiently by simply stimulating peripheral blood mononuclear cells with viral peptide and anti-4-1BB antibody without using antigen-presenting cells. We propose using anti-4-1BB antibody as a novel strategy to generate VSTs for adoptive therapy.


Abdominal Ectopic Pregnancy and Impaired Postnatal Mammary Gland Development, Consistent With Physiologic Agalactia, in a Wild European Rabbit, Oryctolagus cuniculus.
A wild European rabbit, Oryctolagus cuniculus, was diagnosed with an abdominal pregnancy due to the presence of a single abdominal lithopedion attached by a thin fibrovascular stalk to the left uterine horn, which was distorted by the tension of the adhesion. Evidence of mineralized remnants, chronic inflammation, and fibrosis in the left uterine endometrium and myometrium suggested that the lithopedion had arisen as a secondary abdominal pregnancy. The right uterine horn contained two macroscopically normal fetuses. The mammary gland exhibited notably retarded development in relation to the size of the fetuses. Histologically, mammary alveoli lacked evidence of intraluminal secretory product, and ducts lacked prominence and contained clusters of small numbers of macrophages. The doe also exhibited mild granulomatous and heterophilic pneumonia with rare intralesional adiaspores, suggesting infection with Emmonsia spp. as an incidental finding. This case documents secondary abdominal pregnancy in a wild lagomorph not subjected to artificial insemination procedures suggested to increase the occurrence of this condition in farmed rabbits. An abdominal pregnancy is one of a number of factors that should be considered as a potential factor in the etiology of impaired postnatal mammary development or reduced milk yield in a breeding doe, although no causative association is demonstrated in this case. Abdominal ectopic pregnancy is one possible differential diagnosis in the investigation of the presence of a palpable abdominal mass or masses in O. cuniculus.


α-Galactosidase and Sucrose-Kinase Relationships in a Bi-functional AgaSK Enzyme Produced by the Human Gut Symbiont Ruminococcus gnavus E1.
Plant α-galactosides belonging to the raffinose family oligosaccharides (RFOs) and considered as prebiotics, are commonly degraded by α-galactosidases produced by the human gut microbiome. In this environment, the Ruminococcus gnavus E1 symbiont-well-known for various benefit-is able to produce an original RgAgaSK bifunctional enzyme. This enzyme contains an hydrolytic α-galactosidase domain linked to an ATP dependent extra-domain, specifically involved in the α-galactoside hydrolysis and the phosphorylation of the glucose, respectively. However, the multi-modular relationships between both catalytic domains remained hitherto unexplored and has been, consequently, herein investigated. Biochemical characterization of heterologously expressed enzymes either in full-form or in separated domains revealed similar kinetic parameters. These results were supported by molecular modeling studies performed on the whole enzyme in complex with different RFOs. Further enzymatic analysis associated with kinetic degradation of various substrates followed by high pressure anionic exchange chromatography revealed that catalytic efficiency decreased as the number of D-galactosyl moieties branched onto the oligosaccharide increased, suggesting a preference of RgAgaSK for RFO's short chains. A wide prevalence and abundance study on a human metagenomic library showed a high prevalence of the RgAgaSK encoding gene whatever the health status of the individuals. Finally, phylogeny and synteny studies suggested a limited spread by horizontal transfer of the clusters' containing RgAgaSK to only few species of Firmicutes, highlighting the importance of these undispersed tandem activities in the human gut microbiome.


Low-threshold transient calcium current in rat hippocampal lacunosum-moleculare interneurons: kinetics and modulation by neurotransmitters.
Interneurons from the CA1 lacunosum-moleculare (L-M) region were isolated by trypsin-hyaluronidase treatment and mechanical trituration of the L-M. Interneurons isolated in this manner were multipolar with several dendritic processes and could be distinguished from CA1 pyramidal neurons. The properties of a low-threshold transient (LTT) Ca2+ current were investigated using whole-cell voltage-clamp techniques. The activation threshold of the LTT Ca2+ current was -60 mV, and the peak current, 100 +/- 9 pA (mean +/- SEM; n = 15), was observed at -30 mV. Ca2+ was the predominant charge carrier because the current was not affected by tetrodotoxin and was abolished in Ca(2+)-free external solution. Steady state inactivation was observed when the holding potential was positive to -100 mV, and the current was half-inactivated at -84 mV. Complete inactivation occurred at a holding potential of -60 mV. The time-to-peak of the current was highly voltage dependent and ranged from 10 msec at -60 mV to 4 msec at 0 mV. The time constant of inactivation was also voltage dependent and ranged from 27 msec at -60 mV to 12 msec at greater than -30 mV. Recovery from inactivation to 90% of maximum current occurred within 200 msec. L-M interneurons receive synaptic inputs from the septum that release ACh or GABA and from the raphe nuclei that release 5-HT. Carbachol, a nonhydrolyzable cholinergic agonist, and 5-HT quickly and reversibly increased the amplitude of the LTT Ca2+ current. Carbachol's actions were blocked by atropine, indicating that this effect was mediated by muscarinic receptors.(ABSTRACT TRUNCATED AT 250 WORDS)


An application of Organismic Integration Theory to enhance basic psychological needs satisfaction and motivation for rehabilitation in older stroke survivors: A randomized controlled trial study.
Stroke survivors may experience disability and need long-term post-stroke rehabilitation to maintain optimal functioning. However, rehabilitation may not be sufficiently performed due to lack of motivation. This randomized controlled trial aimed to investigate the effectiveness of the Organismic Integration Theory (OIT)-based program for enhancing basic psychological needs satisfaction and motivation for rehabilitation in older stroke survivors. Participants were 38 older stroke survivors randomly assigned to an experimental group (n = 19) receiving the OIT-based program and a control group (n = 19) receiving standard care. Data were collected at baseline, and at 1, 4, and 12 weeks after the program ended. Data analysis showed the significantly higher levels of basic psychological satisfaction and motivation for rehabilitation in participants receiving the OIT-based program than those receiving standard care. The findings support the effectiveness of the OIT-based program in enhancing basic psychological satisfaction and motivation for home rehabilitation of older stroke survivors.


Development of Anthraquinone Derivatives as Ectonucleoside Triphosphate Diphosphohydrolase (NTPDase) Inhibitors With Selectivity for NTPDase2 and NTPDase3.
Ectonucleoside triphosphate diphosphohydrolases (NTPDases) catalyze the hydrolysis of nucleoside tri- and di-phosphates to mono-phosphates. The products are subsequently hydrolyzed by ecto-5'-nucleotidase (ecto-5'-NT) to nucleosides. NTPDase inhibitors have potential as novel drugs, e.g., for the treatment of inflammation, neurodegenerative diseases, and cancer. In this context, a series of anthraquinone derivatives structurally related to the anthraquinone dye reactive blue-2 (RB-2) was synthesized and evaluated as inhibitors of human NTPDases utilizing a malachite green assay. We identified several potent and selective inhibitors of human NTPDase2 and -3. Among the most potent NTPDase2 inhibitors were 1-amino-4-(9-phenanthrylamino)-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (20, PSB-16131, IC50 of 539 nM) and 1-amino-4-(3-chloro-4-phenylsulfanyl)phenylamino-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (48, PSB-2020, IC50 of 551 nM). The most potent NTPDase3 inhibitors were 1-amino-4-[3-(4,6-dichlorotriazin-2-ylamino)-4-sulfophenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (42, PSB-1011, IC50 of 390 nM) and 1-amino-4-(3-carboxy-4-hydroxyphenylamino)-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (33, PSB-2046, IC50 of 723 nM). The best NTPDase2 inhibitor 20 showed a non-competitive inhibition type, while the NTPDase3 inhibitor 42 behaved as a mixed-type inhibitor. These potent compounds were found to be selective vs. other NTPDases. They will be useful tools for studying the roles of NTPDase2 and -3 in physiology and under pathological conditions.


Nivolumab Monotherapy and Nivolumab Plus Ipilimumab in Recurrent Small Cell Lung Cancer: Results From the CheckMate 032 Randomized Cohort.
Nivolumab monotherapy is approved in the United States for third-line or later metastatic small cell lung cancer based on pooled data from nonrandomized and randomized cohorts of the multicenter, open-label, phase 1/2 trial of nivolumab ± ipilimumab (CheckMate 032; NCT01928394). We report updated results, including long-term overall survival (OS), from the randomized cohort. Patients with small cell lung cancer and disease progression after one to two prior chemotherapy regimens were randomized 3:2 to nivolumab 3 mg/kg every 2 weeks or nivolumab 1 mg/kg plus ipilimumab 3 mg/kg every 3 weeks for four cycles followed by nivolumab 3 mg/kg every 2 weeks. Patients were stratified by number of prior chemotherapy regimens and treated until disease progression or unacceptable toxicity. The primary endpoint was objective response rate (ORR) by blinded independent central review. Overall, 147 patients received nivolumab and 96 nivolumab plus ipilimumab. Minimum follow-up for ORR/progression-free survival/safety was 11.9 months (nivolumab) and 11.2 months (nivolumab plus ipilimumab). ORR increased with nivolumab plus ipilimumab (21.9% versus 11.6% with nivolumab; odds ratio: 2.12; 95% confidence interval: 1.06-4.26; p = 0.03). For long-term OS, minimum follow-up was 29.0 months (nivolumab) versus 28.4 months (nivolumab plus ipilimumab); median (95% confidence interval) OS was 5.7 (3.8-7.6) versus 4.7 months (3.1-8.3). Twenty-four-month OS rates were 17.9% (nivolumab) and 16.9% (nivolumab plus ipilimumab). Grade 3 to 4 treatment-related adverse event rates were 12.9% (nivolumab) versus 37.5% (nivolumab plus ipilimumab), and treatment-related deaths were n =1 versus n = 3, respectively. Whereas ORR (primary endpoint) was higher with nivolumab plus ipilimumab versus nivolumab, OS was similar between groups. In each group, OS remained encouraging with long-term follow-up. Toxicities were more common with combination therapy versus nivolumab monotherapy.


Vpu and Tsg101 regulate intracellular targeting of the human immunodeficiency virus type 1 core protein precursor Pr55gag.
Assembly of human immunodeficiency virus type 1 (HIV-1) is directed by the viral core protein Pr55gag. Depending on the cell type, Pr55gag accumulates either at the plasma membrane or on late endosomes/multivesicular bodies. Intracellular localization of Pr55gag determines the site of virus assembly, but molecular mechanisms that define cell surface or endosomal targeting of Pr55gag are poorly characterized. We have analyzed targeting of newly synthesized Pr55gag in HeLa H1 cells by pulse-chase studies and subcellular fractionations. Our results indicated that Pr55gag was inserted into the plasma membrane and, when coexpressed with the viral accessory protein Vpu, Pr55gag remained at the plasma membrane and virions assembled at this site. In contrast, Pr55gag expressed in the absence of Vpu was initially inserted into the plasma membrane, but subsequently endocytosed, and virus assembly was partially shifted to internal membranes. This endocytosis of Pr55gag required the host protein Tsg101. These results identified a previously unknown role for Vpu and Tsg101 as regulators for the endocytic uptake of Pr55gag and suggested that the site of HIV-1 assembly is determined by factors that regulate the endocytosis of Pr55gag.


Determination of a third orthogonal velocity component using two rotationally displaced laser Doppler velocimeter systems.
Transformation equations are derived that may be used to determine a third orthogonal velocity component from measurements made at a common point by two rotationally displaced, two orthogonal component laser Doppler velocimeter (LDV) systems. These equations also may be used to relate velocity measurements made in a particular coordinate system to any other coordinate system. It is shown that the set of smallest angles that may be used to separate the two, two component, LDV systems is very sensitive to the relative magnitudes and directions of the individual velocity components. When the magnitudes of all velocity components are about the same, it is found that the minimum angle of separation is of the same order as the instrumental error of the LDV systems.


Biochemical characterization of the murine activated lymphocyte alloantigen Ly-6E.1 controlled by the Ly-6 locus.
The Ly-6.1 alloantigen defined by monoclonal antibody SK70.94 and expressed predominantly on activated lymphocytes has been immunoprecipitated from lysates of cells biosynthetically radiolabeled with 35S-methionine. On SDS-polyacrylamide gel electrophoresis the reduced antigen appeared as a doublet of 17 and 18 kD. The nonreduced polypeptides had higher mobilities indicating intrachain but not interchain disulfide linkages. The nonreduced form was detectable by immunoperoxidase stain on blots after SDS-PAGE. This showed both polypeptide species contained the antigenic epitope. Labeling in the presence of tunicamycin did not change the apparent m.w. suggesting the absence of N-linked carbohydrate. Pulse-chase data are inconsistent with a strict precursor-product relationship between the two polypeptides and give a half-life for the antigen in 2-day Con A blast cells of about 4 1/2 hr. A highly purified preparation of this antigen displayed a similar electrophoretic pattern and, in the presence of deoxycholate, eluted from a Sephacryl S-200 gel filtration column with a partition coefficient equivalent to that of a 31-kD soluble globular protein. Because of the associated detergent and probable deviation from globularity, this is an over-estimate of the size of the native molecule, and is more consistent with the native molecule being a single polypeptide chain rather than a dimer. Isoelectric focusing of this material showed microheterogeneity with at least five bands between pI 4 and 5. Another monoclonal antibody, HD-42, which had been isolated on the basis of its specificity for a fibrosarcoma antigen subsequently found to be Ly-6 related, precipitated the same polypeptides. Furthermore, no obvious difference was evident between precipitates from Con A-activated lymphocytes and Meth A fibrosarcoma cells.


Selectively Upgrading Lignin Derivatives to Carboxylates through Electrochemical Oxidative C(OH)-C Bond Cleavage by a Mn-Doped Cobalt Oxyhydroxide Catalyst.
Oxidative cleavage of C(OH)-C bonds to afford carboxylates is of significant importance for the petrochemical industry and biomass valorization. Here we report an efficient electrochemical strategy for the selective upgrading of lignin derivatives to carboxylates by a manganese-doped cobalt oxyhydroxide (MnCoOOH) catalyst. A wide range of lignin-derived substrates with C(OH)-C or C(O)-C units undergo efficient cleavage to corresponding carboxylates in excellent yields (80-99 %) and operational stability (200 h). Detailed investigations reveal a tandem oxidation mechanism that base from the electrolyte converts secondary alcohols and their derived ketones to reactive nucleophiles, which are oxidized by electrophilic oxygen species on MnCoOOH from water. As proof of concept, this approach was applied to upgrade lignin derivatives with C(OH)-C or C(O)-C motifs, achieving convergent transformation of lignin-derived mixtures to benzoate and KA oil to adipate with 91.5 % and 64.2 % yields, respectively.


Diet composition and energy balance in humans.
Inpatient metabolic studies of human subjects were performed to obtain data on important nutritional issues. It was shown that wide variations in the ratio of carbohydrate to fat do not alter total 24-h energy need. Studies of the fatty acid composition of plasma low-density lipoproteins during low-fat feeding indicated that there can be considerable lipogenesis from carbohydrate in humans during isoenergetic feeding. The energy cost of this conversion must be small or be counterbalanced by other changes in energy metabolism because measured energy need was unaltered by fat-to-carbohydrate ratios. Energy need was, however, markedly varied by changes in body weight. Subjects at their usual body weights who had experimentally induced increases in body weight became inefficient and required a higher energy intake for weight maintenance. The reverse occurred with a reduction in body weight. The set point at which energy storage is defended is clearly different in obese persons.


Oxidatively stable nanoporous silicon photocathodes with enhanced onset voltage for photoelectrochemical proton reduction.
Stable and high-performance nanoporous "black silicon" photoelectrodes with electrolessly deposited Pt nanoparticle (NP) catalysts are made with two metal-assisted etching steps. Doubly etched samples exhibit an ∼300 mV positive shift in photocurrent onset for photoelectrochemical proton reduction compared to oxide-free planar Si with identical catalysts. We find that the photocurrent onset voltage of black Si photocathodes prepared from single-crystal planar Si wafers by an Ag-assisted etching process increases in oxidative environments (e.g., aqueous electrolyte) owing to a positive flat-band potential shift caused by surface oxidation. However, within 24 h, the surface oxide layer becomes a kinetic barrier to interfacial charge transfer that inhibits proton reduction. To mitigate this issue, we developed a novel second Pt-assisted etch process that buries the Pt NPs deep into the nanoporous Si surface. This second etch shifts the onset voltage positively, from +0.25 V to +0.4 V versus reversible hydrogen electrode, and reduces the charge-transfer resistance with no performance decrease seen for at least two months. PEC performance was stable owing to Pt NP catalysts that were buried deeply in the photoelectrode by the second etch, below a thick surface layer comprised primarily of amorphous SiO2 along with some degree of remaining crystalline Si as observed by scanning and transmission electron micrographs. Electrochemical impedance studies reveal that the second etch leads to a considerably smaller interfacial charge-transfer resistance than samples without the additional etch, suggesting that burying the Pt NPs improves the interfacial contact to the crystalline silicon surface.


Dairy products and plasma cholesterol levels.
Cholesterol synthesized in the body or ingested is an essential lipid component for human survival from our earliest life. Newborns ingest about 3-4 times the amount per body weight through mother's milk compared to the dietary intake of adults. A birth level of 1.7 mmol/L plasma total cholesterol will increase to 4-4.5 mmol/L during the nursing period and continue to increase from adulthood around 40% throughout life. Coronary artery disease and other metabolic disorders are strongly associated with low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol as well as triacylglycerol concentration. Milk fat contains a broad range of fatty acids and some have a negative impact on the cholesterol rich lipoproteins. The saturated fatty acids (SFAs), such as palmitic acid (C16:0), myristic acid (C14:0), and lauric acid (C12:0), increase total plasma cholesterol, especially LDL, and constitute 11.3 g/L of bovine milk, which is 44.8% of total fatty acid in milk fat. Replacement of dairy SFA and trans-fatty acids with polyunsaturated fatty acids decreases plasma cholesterol, especially LDL cholesterol, and is associated with a reduced risk of cardiovascular disease. Available data shows different effects on lipoproteins for different dairy products and there is uncertainty as to the impact a reasonable intake amount of dairy items has on cardiovascular risk. The aim of this review is to elucidate the effect of milk components and dairy products on total cholesterol, LDL, HDL, and the LDL/HDL quotients. Based on eight recent randomized control trials of parallel or cross-over design and recent reviews it can be concluded that replacement of saturated fat mainly (but not exclusively) derived from high-fat dairy products with low-fat dairy products lowers LDL/HDL cholesterol and total/HDL cholesterol ratios. Whey, dairy fractions enriched in polar lipids, and techniques such as fermentation, or fortification of cows feeding can be used to produce dairy products with more beneficial effects on plasma lipid profile.


Co-expression of the SARS-CoV-2 entry molecules ACE2 and TMPRSS2 in human ovaries: Identification of cell types and trends with age.
The high rate of SARS-CoV-2 infection poses a serious threat to public health. Previous studies have suggested that SARS-CoV-2 can infect human ovary, the core organ of the female reproductive system. However, it remains unclear which type of ovarian cells are easily infected by SARS-CoV-2 and whether ovarian infectivity differs from puberty to menopause. In this study, public datasets containing bulk and single-cell RNA-Seq data derived from ovarian tissues were analyzed to demonstrate the mRNA expression and protein distribution of the two key entry receptors for SARS-CoV-2-angiotensin-converting enzyme 2 (ACE2) and type II transmembrane serine protease (TMPRSS2). Furthermore, an immunohistochemical study of ACE2 and TMPRSS2 in human ovaries of different ages was conducted. Differentially expressed gene (DEG) analysis of ovaries of different ages and with varying ovarian reserves was conducted to explore the potential functions of ACE2 and TMPRSS2 in the ovary. The analysis of the public datasets indicated that the co-expression of ACE2 and TMPRSS2 was observed mostly in oocytes and partially in granulosa cells. However, no marked difference was observed in ACE2 or TMPRSS2 expression between young and old ovaries and ovaries with low and high reserves. Correspondingly, ACE2 and TMPRSS2 were detected in the human ovarian cortex and medulla, especially in oocytes of different stages, with no observed variations in their expression level in ovaries of different ages, which was consistent with the results of bioinformatic analyses. Remarkably, DEG analysis showed that a series of viral infection-related pathways were more enriched in ACE2-positive ovarian cells than in ACE2-negative ovarian cells, suggesting that SARS-CoV-2 may potentially target specific ovarian cells and affect ovarian function. However, further fundamental and clinical research is still needed to monitor the process of SARS-CoV-2 entry into ovarian cells and the long-term effects of SARS-CoV-2 infection on the ovarian function in recovered females.


Inhibitory Effects of Polyacetylene Compounds from Panax ginseng on Neurotrophin Receptor-Mediated Hair Growth.
Neurotrophins play an important role in the control of the hair growth cycle. Therefore, neurotrophin receptor antagonists have therapeutic potential for the treatment of hair growth disorders. In this study, we investigated the inhibitory effect of Panax ginseng, a medicinal plant commonly used to treat alopecia, on the binding of neurotrophins to their receptors. In addition, we isolated and characterized the bioactive compounds of P. ginseng extracts. P. ginseng hexane extracts strongly inhibited brain-derived neurotrophic factor (BDNF)-TrkB and β-nerve growth factor (β-NGF)-p75 neurotrophin receptor (p75NTR) binding. Furthermore, we identified the following 6 polyacetylene compounds as the bioactive components in P. ginseng hexane extract: panaxynol (1), panaxydol (2), panaxydol chlorohydrin (3), 1,8-heptadecadiene-4,6-diyne-3,10-diol (4), panaxytriol (5), and dihydropanaxacol (6). In particular, compounds 4, 5, and 6 significantly inhibited BDNF-TrkB binding in a dose-dependent manner. To identify the structural component mediating the inhibitory effect, we investigated the effects of the hydroxyl moiety in these compounds. We found that the inhibitory effect of panaxytriol (5) was strong, whereas the inhibitory effect of Ac-panaxytriol (7) was relatively weak. Our findings suggest that P. ginseng-derived polyacetylenes with a hydroxyl moiety might provide therapeutic benefits to patients with hair growth disorders such as alopecia by inhibiting the binding of neurotrophins to their receptors. Although saponins have been proposed to be the primary mediators of the effects of P. ginseng on hair growth, this study revealed that polyacetylene compounds exert similar effects.


Gingival crevicular fluid alkaline phosphatase activity reflects periodontal healing/recurrent inflammation phases in chronic periodontitis patients.
Roles for host enzymes as diagnostic indicators of periodontal status in gingival crevicular fluid (GCF) have been proposed. One of these host enzymes is alkaline phosphatase (ALP), the GCF activity of which has been associated with periodontal inflammation. Thus, the present study aimed to improve our understanding of how the healing of chronic periodontitis following scaling and root planing (SRP) affects GCF ALP activity after 15 and 60 days. Sixteen systemically healthy subjects (aged 35 to 61 years) with moderate to advanced generalized chronic periodontitis were recruited. In each subject, paired pockets with probing depths (PDs) > or =4 mm that were located in two symmetric quadrants were chosen. These sites were randomized at the split-mouth level, with half receiving SRP treatment and the other half left untreated. Ninety-two pockets were included in the study. Clinical examinations were performed at baseline (prior to SRP) and after 15 and 60 days; information recorded included the presence of plaque, PD, clinical attachment level (CAL), and bleeding on probing. GCF was collected from each pocket included in the study at the three time points. A large and significant decrease in GCF ALP activity was seen 15 days after SRP, concomitant with an improvement in clinical parameters. After 60 days, an increase in GCF ALP activity back to baseline levels was recorded along with further improvements in clinical parameters. Moreover, in the SRP pockets with initial PDs >6 mm, the CAL gains between days 15 and 60 were significantly associated with changes in GCF ALP activity over the same time interval. The decrease in GCF ALP activity at 15 days corresponded to a decrease in clinical signs of inflammation; in contrast, the increase in GCF ALP activity at 60 days seemed to be related to subclinical recurrent inflammation or further healing/remodeling of the periodontal tissue. Therefore, GCF ALP reflects the short-term periodontal healing/recurrent inflammation phases in chronic periodontitis patients.


HIV Stigma, Depressive Symptoms, and Substance Use.
Substance use problems undermine HIV treatment and secondary prevention efforts. Research is needed to better understand predictors of substance use among people living with HIV (PLWH). We examined whether internalized stigma and enacted HIV stigma are associated with three indicators of substance use among PLWH, including numbers of (1) substances used, (2) substances used at moderate to high risk, and (3) times substances were used before sex, through the mediator of depressive symptoms. Participants included 358 PLWH aged 18-35 years from Georgia, United States. At baseline, participants completed measures of internalized and enacted stigma, depressive symptoms, and substance use severity. Substance use was additionally tested with urinalysis. Following baseline, participants reported their use of substances before sex for 28 days through daily text messaging. Data were analyzed using path analysis in R. On average, participants tested positive for 1.24 (range: 0-6) substances used, reported moderate to high risk on 2.01 (range: 0-8) substances, and reported using substances 1.57 (range: 0-20) times before sex over 28 days. Internalized stigma and enacted stigma were associated with greater depressive symptoms, and depressive symptoms were associated with all three indicators of substance use. Moreover, the indirect effects between internalized and enacted stigma with indicators of substance use were significant, suggesting that depressive symptoms partially mediated associations between stigma and substance use. Interventions may be needed to address both stigma and depressive symptoms among PLWH to reduce substance use and support HIV treatment and prevention efforts.


Serratus anterior in vivo contractile force study.
A major limitation of functional muscle transfer for facial and intrinsic hand reanimation is the inability to predict the force that will be generated by the transplanted muscle. The authors studied the contractile force of the slips of the serratus anterior in situ in 10 patients and tested the gracilis muscle in four subjects as a control. Mean contractile force generated by each serratus slip was 0.178 pound (range, 0.019 to 0.797 pound). This compares favorably with the maximum force generated by smiling (0.307 pound). Muscle strength correlated strongly with age (r = -0.805, p = 0.005). The lowest slip generated less force than those above it (0.133 pound versus 0.191 pound); this difference did not reach statistical significance. When the strength of the lowest slip is compared with the more superior slips as a percentage of total force generated by the slips (to compensate for the effect of age on muscle strength), the lowest slip was significantly weaker (18.6 percent of total force versus 25.5 percent of total force, p = 0.013). Mean contractile force generated by the gracilis was 0.963 pound, significantly different from that generated by a serratus anterior slip (p = 0.009). Each serratus slip could potentially be used to generate a separate force vector for facial reanimation. Further separation of the flap along preexisting fascial planes may allow generation of up to 10 independent force vectors, making the serratus anterior muscle flap an attractive option for facial reanimation and possibly intrinsic hand muscle reconstruction.


African-derived genetic polymorphisms in TNFAIP3 mediate risk for autoimmunity.
The TNF alpha-induced protein 3 (TNFAIP3) is an ubiquitin-modifying enzyme and an essential negative regulator of inflammation. Genome-wide association studies have implicated the TNFAIP3 locus in susceptibility to autoimmune disorders in European cohorts, including rheumatoid arthritis, coronary artery disease, psoriasis, celiac disease, type 1 diabetes, inflammatory bowel disease, and systemic lupus erythematosus (SLE). There are two nonsynonymous coding polymorphisms in the deubiquitinating (DUB) domain of TNFAIP3: F127C, which is in high-linkage disequilibrium with reported SLE-risk variants, and A125V, which has not been previously studied. We conducted a case-control study in African-American SLE patients using these coding variants, along with tagging polymorphisms in TNFAIP3, and identified a novel African-derived risk haplotype that is distinct from previously reported risk variants (odds ratio=1.6, p=0.006). In addition, a rare protective haplotype was defined by A125V (odds ratio=0.31, p=0.027). Although A125V was associated with protection from SLE, surprisingly the same allele was associated with increased risk of inflammatory bowel disease. We tested the functional activity of nonsynonymous coding polymorphisms within TNFAIP3, and found that the A125V coding-change variant alters the DUB activity of the protein. Finally, we used computer modeling to depict how the A125V amino acid change in TNFAIP3 may affect the three-dimensional structure of the DUB domain to a greater extent than F127C. This is the first report of an association between TNFAIP3 polymorphisms and autoimmunity in African-Americans.


Alloplastic implantation.
After studying this article, the participant should be able to: 1. Define an alloplastic material and know the differences between an alloplast and other types of implants available for surgical use. 2. Determine the biologic response to alloplastic implantation and the material and host characteristics that contribute to long-term reconstruction success with their use. 3. Review the criteria for choosing a specific alloplastic material for a reconstruction site and the principles of surgical technique for its proper placement. 4. Evaluate the various alloplastic material types that are currently available for surgical use and be able to discuss several physical properties of each as they relate to handling and clinical implantation. 5. Discuss the complication of alloplastic infection, its pathogenesis, preoperative and intraoperative measures for its avoidance, and the postoperative management of its occurrence.


Fluctuations in intracellular pH associated with vasopressin stimulation.
Intracellular pH (pHi) was estimated in paired hemibladders isolated from Dominican toads (Bufo marinus) by the tissue distribution of [14C]5,5'-dimethyl-2,4-oxazolidinedione and [3H]inulin. Tissues were incubated with isotopes for 30 min to correlate changes in pHi with the approximate time of peak vasopressin (VP)-induced water flow. At serosal pH 7.1 in the presence of an osmotic gradient, the intracellular hydrogen ion concentration [( H+]i) after 30 min of VP (20 mU/ml) stimulation was 8.29 +/- 0.23 X 10(-8) M (pHi 7.08) compared with 5.19 +/- 0.46 X 10(-8) M (pHi 7.28) in unstimulated paired controls (n = 5, P less than 0.001). The cyclic AMP (cAMP) analogue 8-(p-chlorophenylthio)-cAMP (10(-5) M) mimicked the VP effects. A similar change was observed at serosal bath pH 8.2, where [H+]i was 1.67 +/- 0.06 X 10(-8) M (pHi 7.78) with VP vs. 1.11 +/- 0.04 X 10(-8) M (pHi 7.95) in matched controls (n = 8, P less than 0.001). In all cases, the hydroosmotic response was associated with a significant decrease in inulin space. When the osmotic gradient was eliminated with Ringer solution or isotonic sorbitol in the mucosal bath, VP produced a smaller decrease in pHi (approximately 0.08 pH units) at both serosal pH. 31P-nuclear magnetic resonance spectra showed a similar downward trend in pHi with cell swelling. When vasopressin was removed from the bath, pHi and inulin space in stimulated hemibladders returned to pretreatment values within 30 min, and the tissues were again capable of a maximum hydroosmotic response if rechallenged with the hormone.(ABSTRACT TRUNCATED AT 250 WORDS)


From microbes to molecules: a review of microbial-driven antioxidant peptide generation.
Oxidative stress, arising from excess reactive oxygen species (ROS) or insufficient antioxidant defenses, can damage cellular components, such as lipids, proteins, and nucleic acids, resulting in cellular dysfunction. The relationship between oxidative stress and various health disorders has prompted investigations into potent antioxidants that counteract ROS's detrimental impacts. In this context, antioxidant peptides, composed of two to twenty amino acids, have emerged as a unique group of antioxidants and have found applications in food, nutraceuticals, and pharmaceuticals. Antioxidant peptides are sourced from natural ingredients, mainly proteins derived from foods like milk, eggs, meat, fish, and plants. These peptides can be freed from their precursor proteins through enzymatic hydrolysis, fermentation, or gastrointestinal digestion. Previously published studies focused on the origin and production methods of antioxidant peptides, describing their structure-activity relationship and the mechanisms of food-derived antioxidant peptides. Yet, the role of microorganisms hasn't been sufficiently explored, even though the production of antioxidant peptides frequently employs a variety of microorganisms, such as bacteria, fungi, and yeasts, which are recognized for producing specific proteases. This review aims to provide a comprehensive overview of microorganisms and their proteases participating in enzymatic hydrolysis and microbial fermentation to produce antioxidant peptides. This review also covers endogenous peptides originating from microorganisms. The information obtained from this review might guide the discovery of novel organisms adept at generating antioxidant peptides.


Walking is more like catching than tapping: gait in the elderly as a complex cognitive task.
Walking is generally viewed as an automated, over-learned, rhythmic motor task and may even be considered the lower-limb analog of rhythmic finger tapping, another automated motor task. Thus, one might hypothesize that walking would be associated with a simple rhythmic task like tapping rather than with a complex motor task like catching. Surprisingly, however, we find that among older adults, routine walking has more in common with complex motor tasks, like catching a moving object, than it does with tapping. Tapping performance, including both the average tapping interval and the variability of tapping interval, was not significantly associated with any gait parameter (gait speed, average stride time and stride time variability). In contrast, catch game performance was significantly associated with measures of walking, suggesting that walking is more like catching than it is like tapping. For example, participants with a higher gait speed tended to have lower times to first move when catching, better catching accuracy, and less catching errors. Stride time variability was significantly associated with each of the measures of catching. Participants with a lower stride time variability (a more steady gait) had better catching accuracy, lower time to first move, fewer direction changes when moving the cursor to catch the falling object, and less catching errors. To understand this association, we compared walking performance to performance on the Stroop test, a classic measure of executive function, and tests of memory. Walking was associated with higher-level cognitive resources, specifically, executive function, but not with memory or cognitive function in general. For example, a lower (better) stride time variability was significantly associated with higher (better) scores on the Stroop test, but not with tests of memory. Similarly, when participants were stratified based on their performance on the Stroop test and tests of memory, stride time variability was dependent on the former, but not the latter. These findings underscore the interconnectedness of gait and cognitive function, indicate that even routine walking is a complex cognitive task that is associated with higher-level cognitive function, and suggest an alternative approach to the treatment of gait and fall risk in the elderly.


Context-dependent regulation of lipid accumulation in adipocytes by a HIF1α-PPARγ feedback network.
Hypoxia-induced upregulation of HIF1α triggers adipose tissue dysfunction and insulin resistance in obese patients. HIF1α closely interacts with PPARγ, the master regulator of adipocyte differentiation and lipid accumulation, but there are conflicting results regarding how this interaction controls the excessive lipid accumulation that drives adipocyte dysfunction. To directly address these conflicts, we established a differentiation system that recapitulated prior seemingly opposing observations made across different experimental settings. Using single-cell imaging and coarse-grained mathematical modeling, we show how HIF1α can both promote and repress lipid accumulation during adipogenesis. Our model predicted and our experiments confirmed that the opposing roles of HIF1α are isolated from each other by the positive-feedback-mediated upregulation of PPARγ that drives adipocyte differentiation. Finally, we identify three factors: strength of the differentiation cue, timing of hypoxic perturbation, and strength of HIF1α expression changes that, when considered together, provide an explanation for many of the previous conflicting reports.


Advanced glycation end-products reduce lipopolysaccharide uptake by macrophages.
Hyperglycaemia provides a suitable environment for infections and the mechanisms of glucose toxicity include the formation of advanced glycation end-products (AGEs), which comprise non-enzymatically glycosylated proteins, lipids, and nucleic acid amino groups. Among AGE-associated phenotypes, glycolaldehyde-derived toxic AGE (AGE-3) is involved in the pathogenesis of diabetic complications. Internalisation of endotoxin by various cell types contributes to innate immune responses against bacterial infection. An endotoxin derived from Gram-negative bacteria, lipopolysaccharide (LPS), was reported to enhance its own uptake by RAW264.7 mouse macrophage-like cells, and an LPS binding protein, CD14, was involved in the LPS uptake. The LPS uptake induced the activation of RAW264.7 leading to the production of chemokine CXC motif ligand (CXCL) 10, which promotes T helper cell type 1 responses. Previously, we reported that AGE-3 was internalised into RAW264.7 cells through scavenger receptor-1 Class A. We hypothesized that AGEs uptake interrupt LPS uptake and impair innate immune response to LPS in RAW264.7 cells. In the present study, we found that AGE-3 attenuated CD14 expression, LPS uptake, and CXCL10 production, which was concentration-dependent, whereas LPS did not affect AGE uptake. AGEs were reported to stimulate the receptor for AGEs and Toll-like receptor 4, which cause inflammatory reactions. We found that inhibitors for RAGE, but not Toll-like receptor 4, restored the AGE-induced suppression of CD14 expression, LPS uptake, and CXCL10 production. These results indicate that the receptor for the AGE-initiated pathway partially impairs the immune response in diabetes patients.


Protein production using the baculovirus-insect cell expression system.
The baculovirus-insect cell expression system is widely used in producing recombinant proteins. This review is focused on the use of this expression system in developing bioprocesses for producing proteins of interest. The issues addressed include: the baculovirus biology and genetic manipulation to improve protein expression and quality; the suppression of proteolysis associated with the viral enzymes; the engineering of the insect cell lines for improved capability in glycosylation and folding of the expressed proteins; the impact of baculovirus on the host cell and its implications for protein production; the effects of the growth medium on metabolism of the host cell; the bioreactors and the associated operational aspects; and downstream processing of the product. All these factors strongly affect the production of recombinant proteins. The current state of knowledge is reviewed.


Developmental study of the anal tonsil in the laboratory shrew, Suncus murinus.
The laboratory shrew, Suncus murinus, which lacks such gut associated lymph organs as the appendix and Peyer's plates, was recently demonstrated (Kubo and Isomura, 1996) to possess a pair of anal tonsils at the end of its rectum. The present paper deals with the development of this lymphoid organ as observed by light and electron microscopy. The anal tonsil was characterized by the initial postnatal development. On neonatal Day 1, a pair of epithelial crypts formed at the dorsal boundary between the anus and the ostium urogenitoanale. On Day 2 after birth, lymphocytes began to accumulate in the subepithelial mesenchymal tissue under the crypt. From Day 3 on, the lymphocytes increased to form a lymph nodule, from which, on Day 5, some lymphocytes began to penetrate into the crypt epithelium. The crypt and the nodule were fused together between Days 6 and 8. A germinal center-like structure was observed on Day 20 after birth. Around Day 40, the invading cells comprised cellular units consisting of large and small lymphocytes and plasma cells. High endothelial venules were observed in the parafollicular area at this time. These findings indicate that the anal tonsil originates from an accumulation of lymphocytes in the mesenchymal tissue close to a particular epithelium of the crypt, presumably in response to antigens in foods; the tonsilar structure is then gradually completed by fusion of the lymphoid and epithelial elements. This paper further reports on an electron microscope finding on Day 8 where the anal tonsillar crypt epithelium was seen to contain some basal-granulated cells of the open type.


Dynamics of suspensions of hydrodynamically structured particles: analytic theory and applications to experiments.
We present an easy-to-use analytic toolbox for the calculation of short-time transport properties of concentrated suspensions of spherical colloidal particles with internal hydrodynamic structure, and direct interactions described by a hard-core or soft Hertz pair potential. The considered dynamic properties include self-diffusion and sedimentation coefficients, the wavenumber-dependent diffusion function determined in dynamic scattering experiments, and the high-frequency shear viscosity. The toolbox is based on the hydrodynamic radius model (HRM) wherein the internal particle structure is mapped on a hydrodynamic radius parameter for unchanged direct interactions, and on an existing simulation data base for solvent-permeable and spherical annulus particles. Useful scaling relations for the diffusion function and self-diffusion coefficient, known to be valid for hard-core interaction, are shown to apply also for soft pair potentials. We further discuss extensions of the toolbox to long-time transport properties including the low-shear zero-frequency viscosity and the long-time self-diffusion coefficient. The versatility of the toolbox is demonstrated by the analysis of a previous light scattering study of suspensions of non-ionic PNiPAM microgels [Eckert et al., J. Chem. Phys., 2008, 129, 124902] in which a detailed theoretical analysis of the dynamic data was left as an open task. By the comparison with Hertz potential based calculations, we show that the experimental data are consistently and accurately described using the Verlet-Weis corrected Percus-Yevick structure factor as input, and for a solvent penetration length equal to three percent of the excluded volume radius. This small amount of solvent permeability of the microgel particles has a significant dynamic effect at larger concentrations.


Spontaneous resolution of occlusive lesions of the carotid artery.
The authors review the literature concerning spontaneous resolution of occlusive lesions involving the carotid artery. They add four cases illustrating three pathological vascular processes that may resolve spontaneously, namely, arterial dissection, atheromatous lesions at the carotid bifurcation, and an arteriopathy involving the intracranial vessels which is poorly understood at this time.


Inosine preserves ATP during ischemia and enhances recovery during reperfusion.
The effects of exogenous inosine (IN) on high-energy phosphate metabolism and function in isolated, working rabbit hearts were monitored with 31P-nuclear magnetic resonance spectroscopy. Dynamic measurements of ATP and phosphocreatine (PCr) were made along with concomitant functional recordings during normal perfusion, global ischemia (IS), and reperfusion (RE). We found that 0.1 mM IN enhanced the rate of pressure development (dP/dt) within the left ventricle by 10 +/- 5% (n = 7). Although IN levels in treated hearts were elevated during normal perfusion, no effect was observed on ATP or PCr levels. However during IS, pretreatment with IN minimized ATP loss for the first 20 min relative to untreated controls (UNT, P less than 0.05). Both IN and UNT hearts that were ischemic for only 13.5 min regained function during a 60-min RE period. However, at the end of IS, IN hearts (n = 8) displayed 88 +/- 10% of the pre-IS ATP levels, whereas UNT hearts (n = 7) retained only 60 +/- 10%. With RE, ATP in IN hearts remained elevated over that of UNT hearts for the entire 60 min. IN treatment also increased the rate of recovery of dP/dt and maintained improved function over 60 min of RE. No correlation was found between post-IS ATP levels and dP/dt values during RE in either IN or UNT hearts. These data indicate that IN was protective against ATP loss during IS and improved functional recovery on RE.


Recurrence of equinus foot deformity in cerebral palsy patients following surgery: a review.
Although equinus deformity in children with cerebral palsy is often corrected with surgery, postsurgical recurrence of the deformity is not uncommon. In order to isolate factors that may be related to its recurrence, 31 studies were evaluated. Data from nine articles indicated that children younger than approximately 7 years of age at the time of surgery had a higher risk of recurrence than children who were older at the time of surgery. Recurrence rates may be understated in studies including less than a minimum of 4-5 years of patient follow-up. Lower recurrence rates are documented in diplegic patients compared with hemiplegic patients, and postoperative casting/splinting is stated, but not documented, to reduce recurrence. One study demonstrated that the use of chemodenervation delayed surgery and by inference theoretically would decrease recurrence after surgical release.


3D models of fungal chromosomes to enhance visual integration of omics data.
The functions of eukaryotic chromosomes and their spatial architecture in the nucleus are reciprocally dependent. Hi-C experiments are routinely used to study chromosome 3D organization by probing chromatin interactions. Standard representation of the data has relied on contact maps that show the frequency of interactions between parts of the genome. In parallel, it has become easier to build 3D models of the entire genome based on the same Hi-C data, and thus benefit from the methodology and visualization tools developed for structural biology. 3D modeling of entire genomes leverages the understanding of their spatial organization. However, this opportunity for original and insightful modeling is underexploited. In this paper, we show how seeing the spatial organization of chromosomes can bring new perspectives to omics data integration. We assembled state-of-the-art tools into a workflow that goes from Hi-C raw data to fully annotated 3D models and we re-analysed public omics datasets available for three fungal species. Besides the well-described properties of the spatial organization of their chromosomes (Rabl conformation, hypercoiling and chromosome territories), our results highlighted (i) in Saccharomyces cerevisiae, the backbones of the cohesin anchor regions, which were aligned all along the chromosomes, (ii) in Schizosaccharomyces pombe, the oscillations of the coiling of chromosome arms throughout the cell cycle and (iii) in Neurospora crassa, the massive relocalization of histone marks in mutants of heterochromatin regulators. 3D modeling of the chromosomes brings new opportunities for visual integration of omics data. This holistic perspective supports intuition and lays the foundation for building new concepts.


Cloning of the heat shock protein 60 gene from the stem borer, Chilo suppressalis, and analysis of expression characteristics under heat stress.
Heat shock protein 60 is an important chaperonin. In this paper, hsp60 of the stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), was cloned by RT-PCR and rapid amplification of cDNA end (RACE) reactions. The full length cDNA of hsp6 degrees Consisted of 2142 bp, with an ORF of 1719 bp, encoding 572 amino acid residues, with a 5'UTR of 158 bp and a 3'UTR of 265 bp. Cluster analysis confirmed that the deduced amino acid sequence shared high identity with the reported sequences from other insects (77%-86%). To investigate whether hsp60 in C. suppressalis responds to thermal stress, the expression levels of hsp60 mRNA in larval haemocytes across temperature gradients from 31 to 39 degrees C were analysed by real-time quantitative PCR. There was no significant difference for hsp60 expression from 28 to 31 degrees C. he temperatures for maximal induction of hsp60 expression in haemocytes was close to 36 degrees C. Hsp60 expression was observed by using flow cytometry. These results revealed that thermal stress significantly induced hsp60 expression and Hsp60 synthesis in larval haemocytes, and the expression profiles of Hsp60 at the mRNA and protein levels were in high agreement with each other from 33 to 39 degrees C.


A Matched Case-Case-Control Study of the Impact of Clinical Outcomes and Risk Factors of Patients with IMP-Type Carbapenemase-Producing Carbapenem-Resistant Enterobacteriaceae in Japan.
IMP-type carbapenemase, found in various Gram-negative bacteria, has been increasingly detected worldwide. We aimed to study the outcomes and risk factors for acquisition of IMP-type carbapenemase-producing carbapenem-resistant Enterobacteriaceae (IMP-CRE), as this has not been evaluated in detail. We conducted a matched case-case-control study of patients from whom IMP-CRE isolates were obtained. All patients who tested positive for IMP-CRE were included; they were matched with patients with carbapenem-susceptible Enterobacteriaceae (CSE) and with controls at a ratio of 1:1:2. The risk factors for acquisition for the CRE and CSE groups and mortality rates, which were calculated using multivariate logistic regression models with weighting according to the inverse probability of propensity scores, were compared. In total, 192 patients (96 patients each in the CRE and CSE groups, with 130 Enterobacter cloacae isolates and 62 Klebsiella sp. isolates) were included. The IMP-11 type was present in 43 patients, IMP-1 in 33, and IMP-60 and IMP-66 in 1 each; 31 patients with CRE (32.3%) and 34 with CSE (35.4%) developed infections. Multivariate analysis identified the following independent risk factors: gastrostomy, history of intravenous therapy or hemodialysis, and previous exposure to broad-spectrum β-lactam antibiotics, including penicillin with β-lactamase inhibitors, cephalosporins, and carbapenems. In propensity score-adjusted analysis, mortality rates for the CRE and CSE groups were similar (15.0% and 19.5%, respectively). We found that IMP-CRE may not contribute to worsened clinical outcomes, compared to CSE, and gastrostomy, previous intravenous therapy, hemodialysis, and broad-spectrum antimicrobial exposure were identified as risk factors for CRE isolation. Fluoroquinolone and aminoglycosides are potentially useful antibiotics for IMP-CRE infections.


Knowledge, attitudes, and practices on climate change and dengue in Lao People's Democratic Republic and Thailand.
Dengue is linked with climate change in tropical and sub-tropical countries including the Lao People's Democratic Republic (Laos) and Thailand. Knowledge about these issues and preventive measures can affect the incidence and outbreak risk of dengue. Therefore, the present study was conducted to determine the knowledge, attitudes, and practices (KAP) among urban and rural communities and government officials about climate change and dengue in Laos and Thailand. A cross-sectional KAP survey about climate change and dengue were conducted in 360 households in Laos (180 urban and 180 rural), 359 households in Thailand (179 urban and 180 rural), and 20 government officials (10 in each country) using structured questionnaires. Data analysis was undertaken using descriptive methods, principal component analysis (PCA), Chi-square test or Fisher's exact test (as appropriate), and logistic regression. Significant differences among the selected communities in both countries were found in terms of household participant's age, level of education, socioeconomic status, attitude level of climate change and KAP level of dengue (P < 0.05; 95% CI). Overall, participants' KAP about climate change and dengue were low except the attitude level for dengue in both countries. The level of awareness among government officials regarding the climatic relationship with dengue was also low. In Lao households, participants' knowledge about climate change and dengue was significantly associated with the level of education and socioeconomic status (SES) (P < 0.01). Their attitudes towards climate change and dengue were associated with educational level and internet use (P < 0.05). Householders' climate change related practices were associated with SES (P < 0.01) and dengue related practices were associated with educational level, SES, previous dengue experience and internet use (P < 0.01). In Thailand, participants' knowledge about climate change was associated with the level of education and SES (P < 0.01). Their attitudes towards climate change were associated with residence status (urban/rural) and internet use (P < 0.05); climate change related practices were associated with educational level and SES (P < 0.05). Dengue related knowledge of participants was associated with SES and previous dengue experience (P < 0.05); participants' dengue related attitudes and practices were associated with educational level (P < 0.01). The findings call for urgently needed integrated awareness programs to increase KAP levels regarding climate change adaptation, mitigation and dengue prevention to improve the health and welfare of people in these two countries, and similar dengue-endemic countries.


Drug-induced nephrotoxicity caused by amphotericin B lipid complex and liposomal amphotericin B: a review and meta-analysis.
Lipid preparations of amphotericin B, commonly used to treat fungal infections, have been demonstrated to have reduced nephrotoxicity compared to conventional amphotericin B. However, to our knowledge, a comprehensive comparison of nephrotoxicity induced by different lipid preparations of amphotericin B has not been performed. We conducted a meta-analysis to evaluate nephrotoxicity associated with amphotericin B lipid complex (ABLC) and liposomal amphotericin B (L-AmB). We searched the PubMed MEDLINE database and abstracts presented at key scientific meetings, and identified 11 studies reported between 1995 and 2008 that compared nephrotoxicity resulting from the use of these agents. Eight of the 11 studies were included in the meta-analysis. The Cochran-Mantel-Haenszel test was used to determine odds ratio (OR) and relative risk (RR), and the Breslow-Day test was used to analyze homogeneity of ORs across different studies. Analysis of all 8 studies (n = 1160) included in the meta-analysis showed an increased probability of nephrotoxicity in patients treated with ABLC versus L-AmB (OR, 1.75; RR, 1.55), but there was a significant lack of homogeneity across these studies (p < 0.001). After excluding the study by Wingard et al, the probability of experiencing nephrotoxicity was more similar between the 2 AmB lipid preparations (OR, 1.31; RR, 1.24; n = 916), particularly when the analysis included only the salvage patient population reported by Hachem et al (OR, 1.12; RR, 1.09; n = 839); the 7 remaining studies were more homogenous by Breslow-Day test (p = 0.054). Our results suggest that nephrotoxicity is generally similar for ABLC and L-AmB in patients receiving antifungal therapy and prophylaxis.


Genetic polymorphisms of the IL6 and NOD2 genes are risk factors for inflammatory reactions in leprosy.
The pathways that trigger exacerbated immune reactions in leprosy could be determined by genetic variations. Here, in a prospective approach, both genetic and non-genetic variables influencing the amount of time before the development of reactional episodes were studied using Kaplan-Meier survival curves, and the genetic effect was estimated by the Cox proportional-hazards regression model. In a sample including 447 leprosy patients, we confirmed that gender (male), and high bacillary clinical forms are risk factors for leprosy reactions. From the 15 single nucleotide polymorphisms (SNPs) at the 8 candidate genes genotyped (TNF/LTA, IFNG, IL10, TLR1, NOD2, SOD2, and IL6) we observed statistically different survival curves for rs751271 at the NOD2 and rs2069845 at the IL6 genes (log-rank p-values = 0.002 and 0.023, respectively), suggesting an influence on the amount of time before developing leprosy reactions. Cox models showed associations between the SNPs rs751271 at NOD2 and rs2069845 at IL6 with leprosy reactions (HRGT = 0.45, p = 0.002; HRAG = 1.88, p = 0.0008, respectively). Finally, IL-6 and IFN-γ levels were confirmed as high, while IL-10 titers were low in the sera of reactional patients. Rs751271-GT genotype-bearing individuals correlated (p = 0.05) with lower levels of IL-6 in sera samples, corroborating the genetic results. Although the experimental size may be considered a limitation of the study, the findings confirm the association of classical variables such as sex and clinical forms with leprosy, demonstrating the consistency of the results. From the results, we conclude that SNPs at the NOD2 and IL6 genes are associated with leprosy reactions as an outcome. NOD2 also has a clear functional pro-inflammatory link that is coherent with the exacerbated responses observed in these patients.


Comparison of physical quality assurance between Scanora 3D and 3D Accuitomo 80 dental CT scanners.
The use of cone beam computed tomography (CBCT) in dentistry has proven to be useful in the diagnosis and treatment planning of several oral and maxillofacial diseases. The quality of the resulting image is dictated by many factors related to the patient, unit, and operator. In this work, two dental CBCT units, namely Scanora 3D and 3D Accuitomo 80, were assessed and compared in terms of quantitative effective dose delivered to specific locations in a dosimetry phantom. Resolution and contrast were evaluated in only 3D Accuitomo 80 using special quality assurance phantoms. Scanora 3D, with less radiation time, showed less dosing values compared to 3D Accuitomo 80 (mean 0.33 mSv, SD±0.16 vs. 0.18 mSv, SD±0.1). Using paired t-test, no significant difference was found in Accuitomo two scan sessions (p>0.05), while it was highly significant in Scanora (p>0.05). The modulation transfer function value (at 2 lp/mm), in both measurements, was found to be 4.4%. The contrast assessment of 3D Accuitomo 80 in the two measurements showed few differences, for example, the grayscale values were the same (SD=0) while the noise level was slightly different (SD=0 and 0.67, respectively). The radiation dose values in these two CBCT units are significantly less than those encountered in systemic CT scans. However, the dose seems to be affected more by changing the field of view rather than the voltage or amperage. The low doses were at the expense of the image quality produced, which was still acceptable. Although the spatial resolution and contrast were inferior to the medical images produced in systemic CT units, the present results recommend adopting CBCTs in maxillofacial imaging because of low radiation dose and adequate image quality.


Bone Mineral Density and Progression of Subclinical Atherosclerosis in African-Americans With Type 2 Diabetes.
Relative to European Americans, calcified atherosclerotic plaque (CP) is less prevalent and severe in African-Americans (AAs). Predictors of progression of CP in the aorta, carotid, and coronary arteries were examined in AAs over a mean 5.3 ± 1.4-year interval. This is the African American-Diabetes Heart Study. A type 2 diabetes (T2D)-affected cohort was included. A total of 300 unrelated AAs with T2D; 50% female, mean age 55 ± 9 years, baseline hemoglobin A1c 8.1 ± 1.8% was included. Glycemic control, renal parameters, vitamin D, and computed tomography-derived measures of adiposity, vascular CP, and volumetric bone mineral density (vBMD) in lumbar and thoracic vertebrae were obtained at baseline and follow-up. CP increased in incidence and quantity/mass in all three vascular beds over the 5-year study (P < .0001). Lower baseline lumbar and thoracic vBMD were associated with progression of abdominal aorta CP (P < .008), but not progression of carotid or coronary artery CP. Lower baseline estimated glomerular filtration rate was associated with progression of carotid artery CP (P = .0004), and higher baseline pericardial adipose volume was associated with progression of coronary artery (P = .001) and aorta (P = .0006) CP independent of body mass index. There was a trend for an inverse relationship between change in thoracic vBMD and change in aortic CP (P = .05). In this longitudinal study, lower baseline thoracic and lumbar vBMD and estimated glomerular filtration rate and higher pericardial adipose volumes were associated with increases in CP in AAs with T2D. Changes in these variables and baseline levels and/or changes in glycemic control, albuminuria, and vitamin D were not significantly associated with progression of CP.


Correction of the Saddle Nose Deformity Using the "Push Up" Technique.
The saddle nose deformity typically occurs after the loss of underlying septal support with loss of projection of the middle nasal vault. This creates a saddling effect as the upper lateral cartilages contract and forms a dip in the profile. The saddle nose deformity can occur from many different etiologies, including septal hematoma, fracture of the septum after trauma, disruption of the dorsal L-strut support after septoplasty, and metabolic disorders resulting in the loss of septal support. In most cases, the nasal bones remain in their native position creating a discrepancy in dorsal height and deformity. Correction of the saddle nose deformity can be performed using one of the two primary approaches. The most basic option is to simply fill the defect with a cartilage onlay graft (solid graft, diced cartilage and fascia, and so on) to create a normal dorsal line. This approach can be effective in many cases. Over time, the onlay graft may descend and create deformity or become visible. The other approach involves opening the middle nasal vault, placing spreader grafts, and reconstituting the dorsal L-strut, performing septal reconstruction or extracorporeal septoplasty. In this paper, we present our technique of repairing the saddle nose deformity using a "push up" method that preserves the middle vault anatomy and reconstitutes the proper projection of the middle-third of the nose.


Re.Ger.O.P.: An Integrated Project for the Recovery of Ancient and Rare Olive Germplasm.
The olive tree is one of the most important economic, cultural, and environmental resources for Italy, in particular for the Apulian region, where it shows a wide diversity. The increasing attention to the continuous loss of plant genetic diversity due to social, economic and climatic changes, has favored a renewed interest in strategies aimed at the recovery and conservation of these genetic resources. In the frame of a project for the valorization of the olive Apulian biodiversity (Re.Ger.O.P. project), 177 minor genotypes were recovered in different territories of the region. They were submitted to morphological, molecular, technological and phytosanitary status analysis in comparison with reference cultivars, then they were propagated and transferred in an ex situ field. All the available information was stored in an internal regional database including photographic documentation and geographic position. The work allowed obtaining information about the genetic diversity of Apulian germplasm, to clarify cases of homonymy and synonymy, to check the sanitary status, and to identify candidate genotypes useful both to set up breeding programs and to enrich the panel of olive cultivars available to farmers for commercial exploitation.


[Purification and properties of cyclic nucleotide phosphodiesterase from uterine tissue].
Cyclic nucleotide phosphodiesterase from calf myometrium has been purified to a homogeneous state for the first time, as can be evidenced from polyacrylamide gel electrophoresis data. The purification procedure included ion-exchange chromatography on DEAE-cellulose, high pressure liquid chromatography on TSK 545 DEAE and gel filtration through Toyopearl HW-55. The molecular mass of the enzyme as determined by gel filtration and polyacrylamide gel electrophoresis is 110 kD. The purified enzyme hydrolyzes cAMP and cGMP with Km = 30 microM and 18 microM, respectively.


The Multiple Mini Interview as a Dental School Admission Tool: Can It Predict Noncognitive Traits Associated with Professional Behaviors?
The aim of this study was to investigate the reliability and validity of the Multiple Mini Interview (MMI) process and to assess its use as an admission tool to predict noncognitive traits associated with professional behavior during patient care in one cohort of dental students at a single U.S. dental school. Data were analyzed for the 95 candidates who matriculated and graduated as part of the 2017 graduating cohort at the University of Michigan School of Dentistry. All MMI interviewees for one cycle of admissions rotated through ten stations: two traditional interview question stations and eight scenario stations measuring domains that included four questions scored on a five-point Likert scale. Generalizability theory analysis showed the MMI to have good reliability (G Coefficient of 0.74). Station reliabilities (Cronbach's alpha) ranged from 0.88 to 0.92. MMI scores showed a positive significant correlation with students' scores on the Dental Admission Test and Perceptual Ability Test, D1 cumulative GPA, and D4 Patient Management grade. MMI scores positively correlated with professional behaviors relating to three domains-openness, conscientiousness, and emotional stability-thus demonstrating good predictive validity for measuring noncognitive traits associated with professionalism. This study found that the MMI was a reliable and valid tool that predicted key behavioral traits associated with professionalism in dental students.


A truncated triangular prism constructed by using imidazole-terpyridine building blocks.
The construction of low-symmetry topological supramolecular structures using bistable building blocks remains challenging. We report an unusual truncated triangular prismatic cage with D3h symmetry using a ligand with both cis- and trans-configurations upon coordination with metal. This work provides new ideas and methods for the future synthesis of low-symmetry topological supramolecules.


Noninvasive measurement of cerebral hemoglobin oxygen saturation using two near infrared spectroscopy approaches.
Spatially resolved spectroscopy (SRS) is a new near infrared spectroscopy (NIRS) method that, using the multi-distance approach, measures local cerebral cortex hemoglobin oxygen saturation [J. Matcher, P. Kirkpatrick, K. Nahid, M. Cope, and D. T. Delpy, Proc. SPIE 2389, 486-495 (1995)]. Using a conventional continuous wave NIRS photometer, cerebral venous oxygen saturation (SvO2) can be calculated from oxyhemoglobin and total hemoglobin rise induced by partial occlusion of jugular vein [C. E. Elwell, S. J. Matcher, L. Tyszczuk, J. H. Meek, and D. T. Delpy, Adv. Exp. Med. Biol. 411, 453-460 (1997)]. The aim of this study was to compare direct measurements of forehead tissue oxygenation index (TOI) with the calculated SvO2 during venous occlusion in 16 adult volunteers using a clinical two-channel SRS oximeter (NIRO-300). Measured TOI and calculated SvO2 values of either right or left forehead did not significantly differ. A good agreement between the two NIRS methods was also demonstrated. On 16 other subjects, no significant differences were found between the right and left forehead TOI values measured simultaneously, and between the TOI values measured by channel 1 or 2 on the same side. The results confirm that cerebral cortex hemoglobin oxygen saturation, measured directly by the SRS method, reflects predominantly the saturation of the intracranial venous compartment of circulation.


[Experimental study of the effect of benzidinsulfon, included in the group of diaminodifenils, with potential carcinogenic effects].
There was performed a study of carcinogenicity of benzidinsulfon (4.4'-diaminodiphenil sulfone) in rats and mice. Experimental animals (99 mice and 99 rats, approximately equally divided by sex) received the drug throughout the life by subcutaneous injections (once a week) or addition to food (5 times a week). A single dose per animal in rats was: subcutaneous administration--50 mg (in females it was reduced due to the toxicity after beginning of the experiment to 25 mg) in 0.5 ml of oil, while feeding--20 mg in 0 5 ml of oil; in mice--respectively 5 mg in 0.2 ml of oil, and 2 mg in 0, 2 ml of oil. The maximum amount of a substance when administered subcutaneously to male rats was 5.65 g, to female rats--2, 68 g, when fed to rats 12.44 g, when injected subcutaneously in mice--380 mg, when fed--737 mg. The survival of experimental animals was significantly reduced as compared to the intact control because of the toxic effect of the drug, preferably chronic nephrosis with nephritic component and secondary nephrosclerosis and as well as miocardiosclerosis and aortic sclerosis. Frequency and timing of detection of tumors in experimental animals was not significantly different from that observed in the control that indicated the absence of carcinogenic features of benzidinsulfon.


The influence of temperature on the threonine and tryptophan requirements of young broiler chicks.
Two experiments were conducted to investigate the influence of a warm environment (35 degrees C) on the threonine and tryptophan requirements of young broiler chicks from 7 to 18 or 21 d of age. Seven hundred forty (experiment 1) and one thousand eight (experiment 2) 1-d-old Cobb x Cobb straight-run broiler chicks were raised in wire-floored battery brooders in moderate temperature rooms (33 to 34 degrees C). For the first 7 d, all chicks were fed a standard corn-soybean-meal-based crumbled starter diet. On d 7, six chicks each (experiment 1) and eight chicks each (experiment 2) were randomly assigned to individual pens. In experiment 1, chicks were fed a corn-peanut meal basal diet supplemented with six levels of threonine (0.630, 0.651, 0.673, 0.715, 0.758, or 0.800% of the diet). In experiment 2, chicks were fed a corn-corn gluten meal based basal diet supplemented with six levels of tryptophan (0.090, 0.115, 0.140, 0.165, 0.190, or 0.215% of the diet). Each dietary treatment was repeated with three pens in each room and three rooms at each temperature. Three rooms were set at a moderate temperature of 25 degrees C, and the other three rooms were set at a warmer temperature of 35 degrees C. Body weight gain, feed consumption, and feed conversion ratio were measured. The threonine requirements of young broiler chicks were 0.733 +/- 0.016% (R2 = 0.59) and 0.752 +/- 0.046% (R2 = 0.25) for body weight gain, and 0.744 +/- 0.016% (R2 = 0.67) and 0.722 +/- 0.016% (R2 = 0.47) for feed conversion ratio at 25 and 35 degrees C, respectively (broken-line linear model). The tryptophan requirements of young broiler chicks were 0.151 +/- 0.004% (R2 = 0.85) and 0.144 +/- 0.003% (R2 = 0.89) for body weight gain, 0.144 +/- 0.003% (R2 = 0.88) and 0.142 +/- 0.003% (R2 = 0.88) for feed consumption, and 0.146 +/- 0.005% (R2 = 0.76) and 0.127 +/- 0.002% (R2 = 0.94) for feed conversion ratio at 25 and 35 degrees C, respectively. On average, the threonine and tryptophan requirements of broiler chicks at 35 degrees C were very similar to those kept at 25 degrees C.


Lactosylceramide mediates tumor necrosis factor-alpha-induced intercellular adhesion molecule-1 (ICAM-1) expression and the adhesion of neutrophil in human umbilical vein endothelial cells.
The endothelial expression of adhesion molecules by proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) has been suggested to contribute to the initiation of atherosclerotic plaque formation. Since lactosylceramide (LacCer) accumulates in large quantities in human atherosclerotic plaque, we have explored its role in TNF-alpha-induced expression of intercellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells and their consequent adhesion to polymorphonuclear leukocytes (PMNs). We found that TNF-alpha increased LacCer synthesis by way of stimulating the activity of UDP-galactose:glucosylceramide beta(1-->4)-galactosyltransferase in a time-dependent fashion. The TNF-alpha-induced expression of ICAM-1 was abrogated by D-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP), an inhibitor of UDP-galactose:glucosylceramide beta(1-->4)-galactosyltransferase. However, the addition of LacCer reversed the D-PDMP effect on TNF-alpha-induced ICAM-1 expression in human umbilical vein endothelial cells. Northern hybridization analysis of mRNA levels and enzyme-linked immunosorbent assays revealed that LacCer (5 microM) specifically stimulated ICAM-1 at both the transcriptional and translational levels. This was accompanied by the adhesion of PMNs, which was visualized by confocal microscopy. Further studies revealed that LacCer stimulated the endogenous generation of superoxide radicals (O-2) about 5-fold compared with the control by specifically activating plasma membrane-associated NADPH-dependent oxidase. This phenomenon was blocked by the antioxidant N-acetyl-L-cysteine, pyrrolidine dithiocarbamate, and the NADPH oxidase inhibitor, diphenylene iodonium. Overexpression of endogeneous CuZn-superoxide dismutase via an adenoviral vector carrying cDNA for CuZn-superoxide dismutase, also inhibited LacCer-induced ICAM-1 expression in endothelial cells. In sum, our findings suggest that LacCer may play the role of a lipid second messenger in TNF-alpha-induced pathogenesis by activating an oxidant-sensitive transcriptional pathway that leads to the adhesion of PMNs to endothelial cells.


Surface chemistry and spectroscopy of UG8 asphaltene Langmuir film, part 2.
While there has been much focus on asphaltenes in toluene, there has been much less focus on asphaltenes in other solvents. It is important to quantify characteristics of asphaltenes in solvents besides toluene in order to better assess their molecular architecture as well as their fundamental aggregation characteristics. The present work focuses on the investigation of UG8 asphaltene Langmuir films at the air-water interface using chloroform as spreading solvent. The results are compared to the results recently obtained using toluene as spreading solvent. Surface pressure-area isotherms and UV-vis spectroscopy indicate that asphaltenes form smaller nanoaggregates in chloroform than in toluene in similar concentration ranges. Still these nanoaggreates share common features with those in toluene. From the surface pressure-area and compression-decompression isotherms, Brewster angle microscopy, and p-polarized infrared reflection-absorption spectroscopy, it was concluded that small size aggregates are spread on the water surface and the compression of the film leads to formation of large aggregates. The films (Langmuir-Schaefer and Langmuir-Blodgett) studied by atomic force microscopy reveal the existence of nanoaggregates spread on the water surface that coexist with large aggregates formed during compression. In addition to these findings, the spreading solvent, chloroform, allows the determination of asphaltene absorption bands using in situ UV-vis spectroscopy at the air-water interface after 15 min waiting time period. The absorbance data carried out after waiting a time period of 1 h shows similar features with the ones carried out after only 15 min; therefore, there is no need to wait 1 h as in the case when toluene is used as spreading solvent. A comparison of the data obtained from chloroform and toluene shows that smaller aggregate sizes are obtained from chloroform as suggested from the surface pressure-area isotherm, in situ UV-vis spectroscopy, and atomic force microscopy. Nevertheless, the similarity of these nanoaggregates in different solvents suggests this formation is a fundamental property of asphaltenes. Moreover, the lack of the isolated absorption band for one-ring aromatics and only a small peak for two-ring aromatics in the UV spectrum of asphaltenes indicate that these groups are not present in asphaltenes in significant quantities.


Dietary sodium intake regulates the ubiquitin-protein ligase nedd4-2 in the renal collecting system.
The activity of the epithelial sodium (Na(+)) channel (ENaC) in the aldosterone-sensitive distal nephron (ASDN) needs to be tightly regulated to match urinary Na(+) excretion with dietary Na(+) intake. The ubiquitin-protein ligase Nedd4-2, which in vitro interacts with ENaC subunits and reduces ENaC cell surface abundance and activity by ubiquitylation of the channel, may participate in the control of ENaC. This study confirms in vivo by reverse-transcriptase-PCR that Nedd4-2 is expressed throughout the nephron and is detectable by immunoblotting in kidney extracts. By immunohistochemistry, Nedd4-2 was found to be strongly expressed in the ASDN, with low staining intensity in the late distal convoluted tubule and early connecting tubule (where apical ENaC is high) and gradually increasing detection levels toward the collecting duct (CD; where apical ENaC is low). Compared with high-Na(+) diet (5% Na(+)), 2 wk of low-Na(+) diet (0.01% Na(+)) drastically reduces Nedd4-2 immunostaining and increases apical ENaC abundance in ASDN. Reduced Nedd4-2 immunostaining is not dependent on increased apical Na(+) entry in the CD, because it is similarly observed in mice with intact and with suppressed apical ENaC activity in the CD. Consistent with a role of mineralocorticoid hormones in the long-term regulation of Nedd4-2, 5-d treatment of cultured CD (mpkCCD(cl4)) cells with 1 microM aldosterone leads to reduction of Nedd4-2 protein expression. It is concluded that Nedd4-2 abundance is regulated by Na(+) diet, by a mechanism that likely involves aldosterone. This regulation may contribute to adaptation of apical ENaC activity to altered Na(+) intake.


Ventilation/perfusion SPECT/CT in patients with pulmonary emphysema. Evaluation of software-based analysing.
The purpose of this study was to evaluate the reproducibility of a new software based analysing system for ventilation/perfusion single-photon emission computed tomography/computed tomography (V/P SPECT/CT) in patients with pulmonary emphysema and to compare it to the visual interpretation. 19 patients (mean age: 68.1 years) with pulmonary emphysema who underwent V/P SPECT/CT were included. Data were analysed by two independent observers in visual interpretation (VI) and by software based analysis system (SBAS). SBAS PMOD version 3.4 (Technologies Ltd, Zurich, Switzerland) was used to assess counts and volume per lung lobe/per lung and to calculate the count density per lung, lobe ratio of counts and ratio of count density. VI was performed using a visual scale to assess the mean counts per lung lobe. Interobserver variability and association for SBAS and VI were analysed using Spearman's rho correlation coefficient. Interobserver agreement correlated highly in perfusion (rho: 0.982, 0.957, 0.90, 0.979) and ventilation (rho: 0.972, 0.924, 0.941, 0.936) for count/count density per lobe and ratio of counts/count density in SBAS. Interobserver agreement correlated clearly for perfusion (rho: 0.655) and weakly for ventilation (rho: 0.458) in VI. SBAS provides more reproducible measures than VI for the relative tracer uptake in V/P SPECT/CTs in patients with pulmonary emphysema. However, SBAS has to be improved for routine clinical use.